HPLC-MS/MS检测双黄连注射液中非法添加13种喹诺酮类药物的研究

建立了高效液相色谱-串联质谱法检测兽用中药制剂双黄连注射液中非法添加13种喹诺酮类药物的方法。样品经0.2%甲酸溶液溶解后稀释,以0.2%甲酸溶液与甲醇作为流动相进行梯度洗脱,采用多反应监测模式进行测定。13种喹诺酮类药物检测限(LOD)为0.1~0.5 mg/m L,定量限(LOQ)为1~5 mg/m L,在5.0~200 ng/m L范围内线性关系良好,相关系数均大于0.99,在5、10、20 mg/m L三个添加浓度下,平均回收率为90.9%~101.9%,相对标准偏差为0.6%~3.8%。     

氟喹诺酮类药物粉剂中非法添加四种喹噁啉类药物的UPLC检测方法研究

建立了超高效液相色谱法测定氟喹诺酮类药物粉剂中非法添加喹乙醇、乙酰甲喹、卡巴氧及喹烯酮的方法。采用purospher RP-18色谱柱(2.1 mm×100 mm,粒径2.0μm)分离四种喹噁啉类药物,以磷酸溶液和甲醇-乙腈(7.5∶7.0)为流动相进行梯度洗脱,流速0.3 m L/min,二极管阵列检测器检测,采集波长范围为200~400 nm,分辨率为1.2 nm,记录光谱图和365 nm波长处的色谱图。结果显示,四种喹噁啉类药物的浓度在0.2~100μg/m L范围内的线性良好,相关系数r均为0.9999,回收率在98.0%~100.2%范围内,RSD在0.27%~0.89%之间,检测限200 mg/kg。本方法快速、准确,可用于氟喹诺酮类粉剂中非法添加喹噁啉类药物的定性和定量检测。     

UPLC-MS/MS法测定3种中兽药制剂中非法添加的阿莫西林和氨苄西林

应用超高效液相色谱-串联质谱技术,建立了3种中兽药制剂中非法添加阿莫西林和氨苄西林的检测方法。样品经50%乙腈水溶液提取、稀释后进行分析。实验采用0.2%甲酸乙腈溶液与0.2%甲酸水溶液作为流动相进行梯度洗脱,经Atlantis T3柱(2.1 mm×150 mm,5μm)分离,通过多反应监测模式进行测定。结果表明,阿莫西林和氨苄西林在1~50 ng/m L的范围内线性关系良好,最低检测限分别为0.5 mg/g和0.2 mg/g。阿莫西林回收率在83.4%~95.8%之间,RSD为2.0%~4.6%;氨苄西林回收率在85.1%~97.2%,RSD为1.7%~4.0%。本方法准确、可靠,可用于中兽药制剂中非法添加阿莫西林和氨苄西林的检测。     

分散固相萃取-高效液相色谱-串联质谱法测定水产品中19种喹诺酮类兽药残留

建立了高效液相色谱-电喷雾串联质谱联用测定水产品中19种喹诺酮类兽药残留的方法。采用酸化乙腈溶液进行提取,用固相填料进行分散固相萃取净化。以甲醇和0.1%甲酸溶液作为流动相,C18作为分析色谱柱,采用梯度洗脱方式进行液相色谱分离,选择离子反应监测模式检测19种喹诺酮类药物,内标方法定量。在2.0~100.0μg/L范围内,19种喹诺酮类药物的线性相关系数均大于0.99。通过添加回收试验,方法的定量限（S/N=10）为1.0μg/kg,3个添加水平中,小龙虾的回收率为70.5%~112.6%,相对标准偏差为3.5%~8.9%,鮰鱼的回收率为70.2%~113.8%,相对标准偏差为4.5%~8.9%。     

超高效液相色谱-串联质谱法测定中兽药散剂中4种喹诺酮药物

为了检测中兽药散剂中喹诺酮类药物的非法添加情况,试验利用超高效液相色谱-串联质谱技术,采用ACQUITY UPLCTM C18色谱柱(2.1 mm×100 mm)进行分离,以乙腈(A)和0.2%甲酸溶液(B)为流动相,梯度洗脱,多反应监测(MRM)扫描,电喷雾电离正离子模式(ESI+),流速为0.4 m L/min,外标法定量。结果表明:4种喹诺酮药物在1~100 ng/m L浓度范围内,药物浓度与吸收峰面积呈现良好的线性关系,相关系数(r)≥0.997。检出限为0.3 ng/m L,定量限为1 ng/m L。该方法可以同时定量检测中兽药散剂中是否添加4种喹诺酮类药物,具有快速、准确、灵敏、成本较低、节省时间、可以应用于高通量的实际检测样品等优点。     

高效液相色谱-串联质谱检测鸡蛋中5种新型喹喏酮类药物

采用超高效液相色谱串联质谱法建立鸡蛋中加雷沙星、曲伐沙星、莫西沙星、吉米沙星和加替沙星5种新型喹诺酮类药物残留的检测方法。样品经过2%甲酸乙腈提取、正己烷分步去脂,氮吹、Oasis HLB柱净化后,经Acquity UPLC BEH Shield RP18(100 mm×2.1 mm,1.7μm)分离,以甲醇和0.1%甲酸水溶液为流动相进行梯度洗脱,多反应监测扫描模式,5种新型喹诺酮类药物能在11 min内完好分离,方法的定量限均低于1.0μg/kg,在5.0~100.0μg/L浓度范围内,5种喹诺酮类药物线性良好,相关系数均在0.992以上;通过2、10、20μg/kg的加标回收试验表明,回收率为63.7%~88.3%,RSD%值为1.36%~11.5%。该方法能满足鸡蛋中新型喹诺酮类药物残留检测要求,对监控养殖过程非法添加隐患具有一定的参考价值。     

超高效液相色谱法同时测定鸡肝中五种氟喹诺酮类药物残留

建立了一种同时测定鸡肝中五种氟喹诺酮类药物残留的超高效液相色谱法(UPLC)。样品经磷酸盐缓冲溶液提取、C18柱净化,采用ACQUITY UPLCTMBEH C18色谱柱(50 mm×2.1mm,1.7μm)分离,以乙腈溶液和0.1%甲酸的水溶液作为流动相进行等度洗脱,荧光检测器检测。五种药物在0.005~0.5μg/mL范围内线性关系良好,相关系数r均大于0.999;以0.05、0.15、0.30μg/g三个浓度水平进行添加回收试验,平均回收率为70.2%~98.2%,相对标准偏差为4.6%~12.1%,方法的检出限(LOD)为0.02μg/g,定量限(LOQ)为0.05μg/g。本方法重现性好、灵敏度高、简单、快捷,适用于鸡肝中五种氟喹诺酮类药物多残留的检测。     

液相色谱-串联质谱法检测牛奶中4种阿维菌素类药物残留量的研究

建立了一种可同时检测牛奶中4种阿维菌素类药物(阿维菌素、伊维菌素、多拉菌素和埃谱利诺菌素)残留的液相色谱-串联质谱方法(LC-MS/MS)。牛奶样品经乙腈提取,高速离心去除蛋白质等杂质,C18柱净化。以0.1%甲酸水溶液和0.1%甲酸乙腈溶液为流动相进行洗脱,流速0.4 m L/min,采用基质匹配标准溶液外标法定量。结果表明:阿维菌素、伊维菌素、多拉菌素和埃谱利诺菌素在0.5~100 ng/m L浓度范围内均呈现良好的线性关系,相关系数(R2)均大于0.996;4种阿维菌素类药物的定量限均为0.5μg/kg。阿维菌素在0.5~5μg/kg添加浓度范围内,伊维菌素在0.5~20μg/kg添加浓度范围内,多拉菌素在0.5~30μg/kg添加浓度范围内,埃谱利诺菌素在0.5~40μg/kg添加浓度范围内,回收率为66.4%~120%;批内与批间相对标准偏差均小于20%。该方法具有简便快速、灵敏度高、定性准确,重复性好等特点,可以满足牛奶中4种阿维菌素类药物残留检测的要求。     

牛奶中七种氟喹诺酮类药物残留检测高效液相色谱-串联质谱法研究

建立了牛奶中氧氟沙星、诺氟沙星、环丙沙星、恩诺沙星、达氟沙星、二氟沙星和沙拉沙星等七种氟喹诺酮类药物(FQs)残留检测的高效液相色谱-串联质谱(HPLC-MS/MS)方法。液相色谱条件为:色谱柱Waters XBridge C18柱(150 mm×2.1 mm,5μm);流动相为0.1%甲酸甲醇溶液 0.1%甲酸水溶液,梯度洗脱;柱温为35℃;流速为0.2 mL/min;进样量为20μL。质谱条件为:电喷雾离子源(ESI ),多反应监测(MRM)方式进行采集;外标法定量。结果表明:七种FQs在5~250 ng/mL浓度范围内呈现良好的线性关系,相关系数R2均大于0.997;方法检测限为2 ng/mL,定量限为5 ng/mL;从10、20和100 ng/mL三个添加浓度检测结果可以看出,方法平均回收率为70.2%~119%,批内RSD为1.8%~16%,批间RSD为9.1%~18%。     

固相萃取/UPLC-MS/MS法检测饲料中18种喹诺酮类药物的研究

为了建立超高效液相色谱-串联质谱法测定饲料中18种喹诺酮类药物的分析方法,以0.1%乙酸乙腈提取饲料样品中的待测物,HLB柱净化,超高效液相色谱-串联质谱法测定,氘代同位素内标法定量。18种喹诺酮类药物在线性范围为0~1.0 mg/L,线性关系良好,相关系数分别在0.9989~0.9999之间,方法的检测限为0.005~0.2 mg/kg。在不同饲料中添加浓度范围为0.1~5.0 mg/kg,平均回收率为43.0%~101%,相对标准偏差小于11.2%。结果表明,该方法灵敏、高效、抗干扰力强,适用于饲料中的18种喹诺酮类药物的测定。     

白术、微米白术和白术多糖对断奶仔猪生长性能和肠道形态及微生态区系的影响

试验选用96头平均体重14.82 kg左右的杜×长×大断奶仔猪,随机分成4组,每组3栏,每栏8头(公母各半)。对照组饲喂基础日粮,试验1、2、3组分别添加1%80目白术、0.2%白术多糖和1%微米白术。试验期30 d。结果表明:在生长性能方面,与对照组相比,1%微米白术添加组可显著提高日增重(P0.05)、降低饲料增重比和腹泻率,而且效果优于1%80目白术组和0.2%白术多糖组,在肠道形态和肠道微生态区系方面,与对照组相比,日粮添加1%80目白术、0.2%白术多糖、1%微米白术均可不同程度的提高十二指肠和空肠的绒毛高度,加深十二指肠和空肠的隐窝深度,并且增加肠道微生态区系的多样性,其中以1%微米白术添加组的效果最佳。     

Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia

Sissay, M.M., Uggla, A. and Waller, P.J., XXXX. Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia. Tropical Animal Health and Production, XXXX. A 2-year abattoir survey was carried out to determine the prevalence, abundance and seasonal incidence of gastro-intestinal (GI) nematodes and trematodes (flukes) of sheep and goats in the semi-arid zone of eastern Ethiopia. During May 2003 to April 2005, viscera including liver, lungs and GI tracts were collected from 655 sheep and 632 goats slaughtered at 4 abattoirs located in the towns of Haramaya, Harar, Dire Dawa and Jijiga in eastern Ethiopia. All animals were raised in the farming areas located within the community boundaries for each town. Collected materials were transported within 24 h to the parasitology laboratory of Haramaya University for immediate processing. Thirteen species belonging to 9 genera of GI nematodes (Haemonchus contortus, Trichostrongylus axei, T. colubriformis, T. vitrinus, Nematodirus filicollis, N. spathiger, Oesophagostomum columbianum, O. venulosum, Strongyloides papillosus, Bunostomum trigonocephalum, Trichuris ovis, Cooperia curticei and Chabertia ovina), and 4 species belonging to 3 genera of trematodes (Fasciola hepatica, F. gigantica, Paramphistomum {Calicohoron} microbothrium and Dicrocoelium dendriticum) were recorded in both sheep and goats. All animals in this investigation were infected with multiple species to varying degrees. The mean burdens of adult nematodes were generally moderate in both sheep and goats and showed patterns of seasonal abundance that corresponded with the bi-modal annual rainfall pattern, with highest burdens around the middle of the rainy season. In both sheep and goats there were significant differences in the mean worm burdens and abundance of the different nematode species between the four geographic locations, with worm burdens in the Haramaya and Harar areas greater than those observed in the Dire Dawa and Jijiga locations. Similar seasonal variations were also observed in the prevalence of flukes. But there were no significant differences in the prevalence of each fluke species between the four locations. Overall, the results showed that Haemonchus, Trichostrongylus, Nematodirus, Oesophagostomum, Fasciola and Paramphistomum species were the most abundant helminth parasites of sheep and goats in eastern Ethiopia.     

Feline intracranial neoplasia: retrospective review of 160 cases (1985-2001)

The purpose of this study was to determine the frequency of different tumor types within a large cohort of cats with intracranial neoplasia and to attempt to correlate signalment, tumor size and location, and survival time for each tumor. Medical records of 160 cats with confirmed intracranial neoplasia evaluated between 1985 and 2001 were reviewed. Parameters evaluated included age, sex, breed, FeLV/FIV status, clinical signs, duration of signs, number of tumors, tumor location(s), imaging results, treatment, survival times, and histopathologic diagnosis. Most of the cats were older (11.3 +/- 3.8 years). Primary tumors accounted for 70.6% of cases. Metastasis and direct extension of secondary tumors accounted for only 5.6 and 3.8% of cases, respectively. Twelve cats (7.5%) had 2 or more discrete tumors of the same type, whereas 16 cats (10.0%) had 2 different types of intracranial tumors. The most common tumor types were meningioma (n = 93, 58.1%), lymphoma (n = 23, 14.4%), pituitary tumors (n = 14, 8.8%), and gliomas (n = 12, 7.5%). The most common neurological signs were altered consciousness (n = 42, 26.2%), circling (n = 36, 22.5%), and seizures (n = 36, 22.5%). Cats without specific neurological signs were common (n = 34, 21.2%). The tumor was considered an incidental finding in 30 (18.8%) cats. In addition to expected relationships (eg, meninges and meningioma, pituitary and pituitary tumors), we found that lesion location was predictive of tumor type with diffuse cerebral or brainstem involvement predictive of lymphoma and third ventricle involvement predictive of meningioma.     

Y‐chromosomal variation of local goat breeds of Turkey close to the domestication centre

Genetic variations in chromosome Y are enabling researchers to identify paternal lineages, which are informative for introgressions and migrations. In this study, the male‐specific region markers, sex‐determining region‐Y (SRY), amelogenin (AMELY) and zinc finger (ZFY) were analysed in seven Turkish native goat breeds, Angora, Kilis, Hair, Honaml?, Norduz, Gürcü and Abaza. A SNP in the ZFY gene defined a new haplotype Y2C. All domestic haplogroups originate from Capra aegagrus, while the finding of Y1A, Y1B, Y2A and Y2C in 32, 4, 126 and 2 Turkish domestic goats, respectively, appears to indicate a predomestic origin of the major haplotypes. The occurrence of four haplotypes in the Hair goat and, in contrast, a frequency of 96% of Y1A in the Kilis breed illustrate that Y‐chromosomal variants have a more breed‐dependent distribution than mitochondrial or autosomal DNA. This probably reflects male founder effects, but a role in adaptation cannot be excluded.     

A brief history of the prevention of infectious diseases by immunisations

Infectious diseases have always been a terrible scourge for humans. The appearance of these plagues, as they were called without distinction, was generally connected to various conditions: asters, climatic changes or religious reasons. The concept of contagious, and then infectious, diseases came slowly. Variolation, i.e. transmission of ‘virulent’ matter to induce a natural disease and the immunity against it, was brought from Constantinople to England by Lady Montague, in 1721. This ‘variolation’ technique was also often performed in veterinary medicine against diseases like sheep-pox or pleuropneumonia. As ‘vaccination’ is the term generally accepted for ‘immunisation’, variolation can be the word designating such a technique. The second period of the history of immunisation began, in 1880, with the studies of Pasteur and his collaborators. A great number of bacterial vaccines were developed: dead, live but attenuated or only parts of pathogens. The viruses were produced in animals, then in eggs and at last, in tissue cultures. Second generation vaccines appeared with genetic engineering: recombinant vaccines, vector vaccines, nucleic acids vaccines, and markers vaccines, among others. These novel technologies can permit the development of new ones and improve the quality of the vaccines already existing.     

Micronutrient Content of Certain Tropical Conventional and Unconventional Feed Resources of Southern India

The ash, silica and certain important micronutrients were estimated in conventional and unconventional feed and fodder resources available in Southern India. Commonly used dry roughages, such as paddy straw, ragi straw, maize kadbi/stalk, jowar kadbi/stalk, bajra stalk and wheat straw, were high in ash (9.9% +/- 0.77%) and silica (6.4% +/- 0.65%) and low in most of the other micronutrients, except iron, with paddy straw containing most silica (>9%). Cultivated non-leguminous (maize, jowar) and improved green crops (hybrid napier, guinea, green panic, NB-21, CO-1) were also high in ash (10.5% +/- 0.60% and 12.5% +/- 0.51%) but were moderate sources of P, Mg and Cu and good sources of Zn (98 +/- 13.8 ppm and 55 +/- 6.7 ppm). Leguminous green fodders (stylosanthus, lucerne, cow pea, soyabean) were excellent sources of Ca (1.9% +/- 0.16%), Mg (0.40% +/- 0.05%), Cu (30 +/- 5.2 ppm), Zn (121 +/- 14.7 ppm) and Fe (1234 +/- 166 ppm) and moderate sources of P. Mixed local grasses and weeds were high in silica (6.9% +/- 1.00%) but were good sources of Cu, Zn and Fe. Cereal grains (maize, wheat, rice, ragi) were low in ash (2.9% +/- 0.33%) and were relatively poor sources of Ca (0.22% +/- 0.03%), Mg (0.19% +/- 0.03%) and Cu (13 +/- 3.1 ppm). Pulses were low to medium sources of most minerals and good sources of Fe (1230 +/- 293 ppm). Oil seed cake/extractions (groundnut cake, cotton seed cake, soyabean meal, sunflower cake, safflower cake) and cereal by-products (rice polish, rice bran, wheat bran) were excellent sources of P (1.1% +/- 0.47% and 2.3% +/- 0.19%) and good sources of Zn (65 +/- 3.9 ppm and 66 +/- 10.7 ppm) and Fe (938 +/- 130 ppm and 662 +/- 126 ppm). Among the unconventional feeds screened, orange peel, sunflower heads, meat meal, rubber seed cake, spirulina algae and sea weeds contained plentiful Ca, Zn and Fe: tree leaves/top feeds (mulberry, erythrina, glyricidia, banana, subabul, groundnut haulms) were excellent sources of Ca (1.5% +/- 0.13%), Zn (120 +/- 22.9 ppm) and Fe (1033 +/- 133 ppm) but relatively poor sources of P. Soyabean husk, cocoa seed husk, rubber seed cake and meat meal were moderate to good sources of P (1.0% and 0.33%). The high Zn and Fe values of most feeds/fodders were probably due to soil contamination. This account of the micronutrient content of feed/fodder resources should help in strategic supplementation intended to alleviate local deficiencies.     

Bovine Mastitis in Selected Areas of Southern Ethiopia

A study on bovine mastitis, designed to determine the causal agents, prevalence of infection and impact of risk factors in three cattle breeds, was conducted in selected areas of southern Ethiopia. A total of 307 lactating and non-lactating cows, of which 162 were indigenous Zebu, 85 Jersey and 60 Holstein-Friesian, were examined by clinical examination and the California mastitis (CMT) test. Of these, 40.4% were positive by CMT and bacteriology for clinical or subclinical mastitis, with prevalence rates of 37.1% and 62.9%, respectively. Out of 1133 quarters examined, 212 (18.7%) were found to be infected, 83 (39.2%) clinically and 129 (60.8%) subclinically. The prevalence of mastitis was significantly higher in Holstein-Friesian than in indigenous Zebu, in non-lactating cows than in lactating cows, in the early lactation stage than in the mid-lactation stage, in cows with lesions and/or tick infestation on skin of udder and/or teats than in cows without this factor, and in the wet season than in the dry season. Mastitis increased with parity number (R = 0.9). Of 248 CMT and clinically positive udder quarter samples analysed microbiologically, 212 were culturally positive for known mastitis pathogens and 36 were negative. Of the 199 positive samples, Staphylococcus accounted for 39.2%, Streptococcus for 23.6%, coliforms for 14.1%, Micrococcus and Bacillus species for 8.0% each and Actinomyces or Arcanobacterium (Corynebacterium) for 7.0%. It was concluded that there was a high prevalence of clinical and subclinical mastitis, mainly caused by Staphylococcus aureus, Streptococcus agalactiae and Escherichia coli, in this study area.     

不同日粮氮水平对山羊氮代谢和微生物蛋白质合成的影响

本试验旨在探讨不同日粮氮水平对内蒙古白绒山羊氮代谢和微生物蛋白质合成(MCP)的影响,以达到提高氮利用率,减少环境污染和饲料资源的浪费。试验选用9只体况良好、体重为(43.83±2.95)kg,装有瘤胃瘘管的内蒙古白绒山羊,按体重随机分为3组,每组3只。日粮分为低氮7.5%、中氮10.5%、高氮13.5%3个氮水平。试验使用常规试验方法分析测定粪尿血液等生化指标,通过全收尿法和尿嘌呤衍生物法估测MCP。结果表明:随着氮水平增加,氨氮(NH3-N)浓度、氮摄入量、尿氮排出、尿中尿素氮(UUN)和MCP显著增加(P0.05),干物质采食量(DMI)显著降低(P0.01);沉积氮占总摄入氮的比例在中氮组最高,MCP在高氮组最高。综上所述,适当降低日粮氮水平可减少粪尿氮排放,提高反刍动物氮利用率。     

Microsporidial keratopathy in two dogs

A microsporidial keratopathy is described in two dogs. Both dogs presented with a unilateral stromal keratopathy characterized by multifocal coalescing opacities, and the diagnosis was made on histopathologic examination of keratectomy specimens. Transmission electron microscopy (TEM) on formalin‐fixed, paraffin‐embedded corneal tissue was performed in one dog, and the morphologic features were consistent with Nosema species infection. Both dogs were initially diagnosed and treated by superficial keratectomy. One dog received additional antifungal medication and underwent a penetrating keratoplasty following local recurrence two years later. No other systemic lesions attributable to the microsporidial infection were identified clinically. The clinical and diagnostic pathology findings, treatment, and follow‐up are discussed.